scholarly journals Inhibition of interleukin 1-stimulated cartilage proteoglycan degradation by a lipophilic inactivator of cysteine endopeptidases

1992 ◽  
Vol 281 (1) ◽  
pp. 175-177 ◽  
Author(s):  
D J Buttle ◽  
J Saklatvala ◽  
M Tamai ◽  
A J Barrett
Keyword(s):  
Interleukin 1 ◽  
Test System ◽  
Significant Inhibition ◽  
Detectable Effect ◽  
Inhibitory Effects ◽  
Proteoglycan Degradation ◽  
Cysteine Endopeptidases ◽  
Cartilage Breakdown ◽  
Pass Through ◽  
Proteoglycan Release

Inactivators of cysteine endopeptidases were tested as inhibitors of the cytokine-stimulated release of proteoglycan from cartilage. The test system consisted of bovine nasal septum cartilage maintained in organ culture, and the stimulus was provided by recombinant human interleukin 1 alpha. L-3-Carboxy-2,3-trans-epoxypropionyl-leucylamido-(4-guanidin o)butane (E64) and L-3-carboxy-2,3-trans-epoxypropionyl-leucylamido-(3-methyl)b utane (Ep475) showed no inhibition at concentrations up to 100 microM. In contrast, trans-epoxysuccinyl-leucylamido-(3-methyl)butane ethyl ester (Ep453), a ‘prodrug’ of Ep475, was an effective inhibitor. The LL-, LD- and DL-isomers gave significant inhibition at 10 microM, and the DD-isomer was inhibitory at 100 microM. None of the isomers had any detectable effect on protein synthesis or glycolysis, and their inhibitory effects were reversible. Iodoacetate inhibited proteoglycan release by a general toxic effect. Our results suggest that cysteine endopeptidase(s) play a part in cytokine-stimulated cartilage breakdown, but that effective inhibitors must pass through membranes.

scholarly journals Lysosomal cysteine endopeptidases mediate interleukin 1-stimulated cartilage proteoglycan degradation

1992 ◽  
Vol 287 (2) ◽  
pp. 657-661 ◽  
Author(s):  
D J Buttle ◽  
J Saklatvala
Keyword(s):  
Interleukin 1 ◽  
Active Form ◽  
Growth Factor Receptor ◽  
Test System ◽  
Significant Inhibition ◽  
Cartilage Explants ◽  
Cartilage Proteoglycan ◽  
Proteoglycan Degradation ◽  
Cysteine Endopeptidases ◽  
Proteoglycan Release

The peptidyl diazomethane inactivator of cysteine endopeptidases, benzyloxycarbonyl-Tyr-Ala-CHN2, was tested as an inhibitor of interleukin 1 alpha-stimulated release of proteoglycan from bovine nasal septum cartilage explants. Like the previously tested epoxidyl peptide proinhibitor trans-epoxysuccinyl-leucylamido-(3-methyl)butane ethyl ester, it proved to be an effective inhibitor of proteoglycan release from cartilage, with significant inhibition at a concentration of 1 microM. The inhibition did not seem to be due to a general toxic effect. The rates of inactivation of the bovine cysteine endopeptidases by the peptidyl diazomethane, the epoxidyl peptide proinhibitor and its active form were determined. Benzyloxycarbonyl-Tyr-Ala-CHN2 proved to be a rapid inactivator of cathepsins L, S and B, but reacted much more slowly with cathepsin H and calpain. Thus it would appear that the latter two enzymes are not implicated in proteoglycan release in our test system. The peptidyl diazomethane and epoxidyl peptide proinhibitor (above) were also tested for their effects on three other interleukin 1-mediated cellular events, namely epidermal growth factor receptor transmodulation, and interleukin 6 and prostaglandin E2 production. In all cases the inactivators did not interfere with the response to interleukin 1 in human gingival fibroblasts. We conclude that one or more of the lysosomal cysteine endopeptidases cathepsins B, L and S mediate interleukin 1-stimulated cartilage proteoglycan degradation without affecting signal transduction.

Role of Receptor Binding and Gene Transcription for Both of Stimulatory and Inhibitory Effects of Interleukin-1 In Pancreatic β-Cells

Autoimmunity ◽  
1992 ◽  
Vol 12 (2) ◽  
pp. 127-133 ◽  
Author(s):  
Décio L. Eizirik ◽  
Daniel E. Tracey ◽  
Klaus Bendtzen ◽  
Stellan Sandler
Keyword(s):  
Gene Transcription ◽  
Receptor Binding ◽  
Interleukin 1 ◽  
Inhibitory Effects ◽  
Β Cells ◽  
Pancreatic Β Cells

cAMP inhibition of interleukin-1-induced interleukin-6 production by human lung fibroblasts

1993 ◽  
Vol 264 (3) ◽  
pp. L253-L260 ◽  
Author(s):  
R. J. Zitnik ◽  
T. Zheng ◽  
J. A. Elias
Keyword(s):  
Human Lung ◽  
Interleukin 1 ◽  
Cell Number ◽  
Lung Fibroblasts ◽  
Intracellular Camp ◽  
Inhibitory Effects ◽  
Mrna Accumulation ◽  
Human Lung Fibroblasts ◽  
Dose Dependent ◽  
Necrosis Factor

We characterized the effects of agents that alter intracellular adenosine 3',5'-cyclic monophosphate (cAMP) on the interleukin (IL)-6 production of human lung fibroblasts. Unstimulated fibroblasts did not produce significant amounts of IL-6. Recombinant (r) tumor necrosis factor (TNF) weakly stimulated, recombinant interleukin-1-alpha (rIL-1 alpha) strongly stimulated, and rIL-1 alpha and rTNF in combination synergistically augmented fibroblast IL-6 production. Prostaglandin (PG)E1, forskolin, dibutyryl cAMP (DBcAMP), 3-isobutyl-1-methylxanthine (IBMX), and cholera toxin did not cause a detectable alteration in the IL-6 production of unstimulated fibroblasts. However, these agents inhibited the IL-6 production of rIL-1 and rIL-1 plus rTNF-stimulated cells. These effects were dose dependent with a concentration of 2 x 10(-9) M PGE1, 5 x 10(-6) M forskolin, 5 x 10(-4) M DBcAMP, and 1 x 10(-3) M IBMX decreasing rIL-1 alpha (2.5 ng/ml)-induced IL-6 production by approximately 50%. The inhibitory effects of these agents, correlated with their ability to induce fibroblast cAMP accumulation, could not be explained by alterations in cell number or viability and were appreciable even when cAMP modifiers were added to fibroblast culture, 1 h after rIL-1. They were also at least partly specific for rIL-1, since these agents increased the IL-6 production of rTNF-stimulated cells. These cAMP-induced alterations in IL-6 production were associated with corresponding alterations in IL-6 mRNA accumulation. Nuclear run-on analysis demonstrated that the inhibitory effects of PGE1 were associated with a comparable decrease in IL-6 transcription. Agents that increase the levels of intracellular cAMP inhibit rIL-1-induced IL-6 by human lung fibroblasts.

Effects of prostaglandins E1, E2, and F2alpha on the growth of leukaemia cells in culture

1976 ◽  
Vol 20 (1) ◽  
pp. 199-206
Author(s):  
T.J. Yang ◽  
J.B. Dale ◽  
R. Machanoff
Keyword(s):  
Tritiated Thymidine ◽  
Inhibitory Effect ◽  
Soft Agar ◽  
Significant Inhibition ◽  
Leucine Incorporation ◽  
Inhibitory Effects ◽  
Uridine Incorporation ◽  
Cells In Culture ◽  
Mouse Leukaemia

Prostaglandins E1, E2, and F2alpha (PGE1, PGE2, and PGF2alpha) were shown to inhibit the growth of mouse leukaemia lymphoblasts L5178Y in culture. The effects of PGE1 and PGE2 were greater than that of PGF2alpha. PGE1 and PGE2, at the concentration of 100 mug per ml showed significant inhibitory effects on the rates of incorporation of tritiated thymidine, uridine and leucine. At concentrations of 50 and 25 mug per ml, there was significant inhibition of thymidine and uridine incorporation, but not of leucine, PGF2alpha showed significant inhibition of thymidine and uridine incorporation but not leucine incorporation, in all 3 concentrations studied (100, 50, and 25 mug/ml). The ability of the cells to form colonies in soft agar was significantly inhibited by PGE1 and PGE2 at concentrations as low as 1–8 mug/ml. For F2alpha, however, a concentration as high as 56mug/ml was required to show inhibitory effect, but at 1–8 mug/ml it was found to be stimulatory.

scholarly journals Differential regulation by cytokines of constitutive and stimulated secretion of von Willebrand factor from endothelial cells

Blood ◽  
1990 ◽  
Vol 75 (3) ◽  
pp. 688-695 ◽  
Author(s):  
EM Paleolog ◽  
DC Crossman ◽  
JH McVey ◽  
JD Pearson
Keyword(s):  
Endothelial Cells ◽  
Von Willebrand Factor ◽  
Messenger Rna ◽  
Cell Injury ◽  
Interleukin 1 ◽  
Tnf Alpha ◽  
Detectable Effect ◽  
Dependent Manner ◽  
Von Willebrand ◽  
Willebrand Factor

Abstract We examined the effect of cytokines on basal and agonist-stimulated release of von Willebrand factor (vWf) by human endothelial cells. Treatment of endothelial cells for up to 48 hours with human recombinant or purified interleukin 1 (IL-1) or human recombinant tumor necrosis factor-alpha (TNF-alpha) did not significantly affect constitutive secretion of vWf or intracellular levels of vWf, although basal prostacyclin (PGI2) production was markedly enhanced. In contrast, both IL-1 and TNF-alpha modulated vWf release in response to thrombin or phorbol ester. Pretreatment of endothelial cells for 2 hours with either cytokine enhanced by up to threefold the stimulatory effect of a subsequent 60-minute exposure to thrombin. Addition of cycloheximide (5 micrograms/mL) during the preincubation abolished this enhancement. Moreover, if the cytokine pretreatment time was extended to 24 hours, agonist-stimulated vWf release was significantly suppressed. Cytokine treatment for 2 or 24 hours had no detectable effect on levels of vWf messenger RNA. The effects of cytokines were not the result of contamination with bacterial lipopolysaccharide and were not attributable to endothelial cell injury. These results show that cytokines have little or no direct effect on vWf release from endothelial cells but can significantly modulate its acute release in response to other stimuli in a complex time- and dose-dependent manner.

Differential effect of basolateral and apical adenosine on AVP-stimulated cAMP formation in primary culture of IMCD

1992 ◽  
Vol 263 (2) ◽  
pp. F268-F276 ◽  
Author(s):  
Y. Yagil
Keyword(s):  
Arginine Vasopressin ◽  
Collecting Duct ◽  
Inhibitory Effect ◽  
Detectable Effect ◽  
Inhibitory Effects ◽  
Tubule Cell ◽  
Adenosine Antagonist ◽  
A1 Receptor ◽  
Medullary Collecting Duct ◽  
Camp Formation

It has been recently established that adenosine interferes with the ability of arginine vasopressin (AVP) to generate adenosine 3',5'-cyclic monophosphate (cAMP) in inner medullary collecting duct (IMCD) cells in culture. The aim of the current study was to determine whether this interaction of adenosine with AVP is mediated by adenosine from the basolateral (B) and/or the apical (A) surface of the tubule cell. Cells from rat IMCD were grown to confluence in monolayers on porous filters. Adenosine (5 x 10(-8)-10(-4) M) applied to the B or A surface of the cell had no detectable effect on basal cAMP formation. AVP, 10(-9)-10(-6) M, increased cAMP formation from both B and A surfaces of the cell. When AVP was applied to the B surface, 10(-6) M adenosine inhibited AVP-stimulated cAMP formation from the B side only, whereas adenosine at 10(-4) M inhibited cAMP formation from both B and A sides. The inhibitory effect of adenosine was reproduced with N6-cyclohexyladenosine (CHA) from both B and A surfaces. 5'-(N-ethylcarboxamido)adenosine (NECA) and 2',5'-dideoxyadenosine (DDA) inhibited cAMP formation from the B surface only. When AVP wasapplied to the A surface, the inhibitory effects of adenosine were the same as when AVP was applied to the B surface; CHA, NECA, and DDA inhibited AVP-stimulated cAMP formation from both the B and A surfaces. 1,3-Dipropyl-8-cyclopentylxanthine (DPCPX), an adenosine antagonist with selectivity for the A1 receptor, prevented the inhibitory effects of adenosine, CHA, and NECA on AVP-stimulated cAMP formation.(ABSTRACT TRUNCATED AT 250 WORDS)

Inhibition of mouse placental lactogen-II release from placental cells by interleukin-1 after mid-pregnancy

1995 ◽  
Vol 147 (3) ◽  
pp. 423-429 ◽  
Author(s):  
M Yamaguchi ◽  
M Sakata ◽  
K Ogura ◽  
K Adachi ◽  
A Mammoto ◽  
...  
Keyword(s):  
Blot Analysis ◽  
Interleukin 1 ◽  
Placental Lactogen ◽  
Inhibitory Effects ◽  
Gm Csf ◽  
Placental Cells ◽  
Steady State Levels ◽  
Combined Action ◽  
Factor Α

Abstract The effects of interleukin (IL)-1 and granulocytemacrophage colony stimulating factor (GM-CSF), which are present in the mouse placenta, on the secretion of mouse placental lactogen (mPL)-1 and mPL-II by placental cells were tested in vitro. IL-lα and IL-1β, 2·5 nmol/l each, significantly inhibited mPL-II secretion by cells from days 9 and 12 of pregnancy, but did not affect mPL-II secretion by cells from day 7 of pregnancy or mPL-I secretion by cells from days 7, 9 or 12 of pregnancy. GM-CSF had no effect on mPL-I and mPL-II secretion by cells from days 7, 9 or 12 of pregnancy. The inhibitory effects of IL-1α and IL-1β on mPL-II secretion were completely eliminated by the addition of antibodies to IL-1α and IL-1β respectively. Western blot analysis for mPL-II indicated that IL-1α significantly reduced the intensity of the mPL-II band. Steady-state levels of mPL-II mRNA, assessed by Northern blot analysis, were reduced by incubation of placental cells from day 12 of pregnancy with 2·5 nmol/l IL-1α for 5 days. Co-incubation of 0·25 pmol/l IL-1α, 25 pmol/l IL-6, and 25 pmol/l tumor necrosis factor-α, each of which did not significantly inhibit mPL-II secretion by itself, together inhibited mPL-II secretion. These results suggest that IL-1, but not GM-CSF, is a potent inhibitor of mPL-II secretion after mid-pregnancy, and that the combined action of cytokines can inhibit mPL-II secretion. Journal of Endocrinology (1995) 147, 423–429

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